biosafety in the tb laboratory presented by peggy coulter with (patient safety monitoring in...
TRANSCRIPT
Biosafety in the TB Laboratory
Presented by Peggy Coulter
with(Patient Safety Monitoring in International Laboratories)
Prepared for the ACTG Conference24 June 2009
Purpose of TrainingGCLP Standards:
DAIDS supported clinical trials and studies involving human subjects must ensure
compliance with federal regulations including procedures to protect the safety of all
participants.Safety of laboratory employees must be a top
priority for all lab facilities.
Abbreviations
• BMBL- Biosafety in Microbiological and Biomedical Laboratories (CDC)
• LBM- Laboratory Biosafety Manual (WHO)• BSC- Biosafety Cabinet• LAI- Laboratory Associated Infection• TB- for Mycobacteriology testing
Objectives: At the end of this training you will be able to:
• Locate resources for biosafety guidelines;• Describe the elements of biosafety;• Identify standard and special practices in
biosafety;• Select and use appropriate biosafety control
measures;• Conduct an internal risk assessment and self
inspection of the laboratory.
Training Topics• Principles of Biosafety• Standard Microbiological Practices• Special Practices• Personal Protective Equipment• Containment Equipment• Laboratory Facilities• Safety Practices• Risk Assessment and Self Inspection
Resources for Principles of Biosafety
• Biosafety in Microbiological and Biomedical Laboratories (BMBL) from CDC at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/BMBL_5th_Edition.pdf
• Laboratory Biosafety Manual from W.H.O. at http://www.who.int/csr/resources/publications/biosafety/Biosafety7.pdf
Introduction to BiosafetyAll laboratory facilities must follow “Standard or Basic Precautions”.
Biosafety guidelines evolved from the microbiological and biomedical community to reduce laboratory
associated infections (LIA’s) and to protect the public health and environment.
Principles of Biosafety
Two basic elements:
• Containment• Risk Assessment
Routes of transmission for LAI’s
• Direct skin, eye, or mucosal membrane;• Parenteral inoculation by a contaminated
sharp or needle;• Ingestion of liquid suspension;• Inhalation of infective aerosols.
Risk Assessment
• Pathogenicity of the microorganism• Prevalence of tuberculosis and rate of MDR• Types of testing performed or referred• Volume of tests• Personnel expertise and attitude• Facilities and equipment
WHO Risk
Group Risk Microorganism
1No or low individual
and community risk.
A microorganism that is unlikely to cause human or animal disease.
2Moderate individual risk, low community
risk.
A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the
community, livestock or the environment. Laboratory exposures may cause serious infection but effective
treatment and preventative measures are available and the risk of spread of infection is limited.
3 High individual risk, low community risk.
A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected
individual to another. Effective treatment and preventative measures are available.
4 High individual and community risk.
A pathogen that usually causes serious human or animal disease and that can be readily transmitted from one individual to another, directly or indirectly. Effective
treatment and preventative measures are not usually available.
Levels of Biosafety, and Testing Levels
Testing Level
Biosafety Level(BSL) Activity
One 2•Collect clinical specimens•Transport specimens to a higher level testing laboratory•May prepare and examine smears of killed tubercle bacilli
Two 3
•Level One activities •Process specimens for microscopy and culture•Identify M. tuberculosis•Perform DST on M. tuberculosis
Three 3
•Level One and Two activities•Identify all Mycobacterium species from clinical specimens•Perform DST against all mycobacteria•Conduct research and provide training to other laboratories
Basic Microbiology Practices
• Policies and access • Safety practices
• Decontamination and Waste• Training
Specimens
Photo of washroom
Handling of Specimens
• Collection• Transportation• Receipt of incoming
specimens• Opening packages
Special Practicesenhance worker safety, provide
environmental protection and address the risk of handling agents
requiring increasing levels of containment.
Reminder: BSL-3 practices should be used whenever M.tuberculosis is handled even if the physical
facilities are a BSL-2.
All persons entering the laboratory must be advised of the potential hazards and meet specific entry/exit requirements.
Animals and plants not associated with the work being performed must not be permitted in the laboratory.
Laboratory personnel must be provided medical surveillance and offered appropriate immunizations for agents handled or potentially
present in the laboratory.
A laboratory-specific biosafety manual must be prepared, adopted as policy and made available and accessible to
the laboratory staff.
This is a written plan thatdefines safe lab practices, spill and emergency
procedures
The laboratory supervisor must ensure that the laboratory personnel
demonstrate proficiency in standard and special microbiological practices
before working in the mycobacteriology lab.
Personal Protective Equipment (PPE)
• Gowns, lab coats• Gloves
• Respirators, masks, goggles, glasses
• Shoe cover, boots
Lab coats vs. Gowns
Gloves
Approved TypeCorrect Size
DonningProper Use
Removal
Shoes and Covers
Open-toed footwear is not appropriate in the laboratory.
Respirator program implemented by the laboratory's safety officer or person designated to perform
this task and should include written procedures concerning how to:
a)select the appropriate respirator, b)conduct fit-testing, and
c)train personnel on the use, fit checking, and storage of the respirator.
Correct Type and Fit of Respirators
Dust and Paint FumesDust
N-95
Surgical Mask
PAPR
Safety Equipment
• Needle locking syringes• Centrifuge safety carriers• Microburners• Biosafety Cabinets (BSC)
Centrifuge Safety
Biosafety Cabinets (BSC)All procedures involving the manipulation
of infectious materials must be conducted within a BSC, or other physical containment devices. No work with open vessels is conducted on the bench. When a procedure cannot be performed within a BSC, a combination of personal protective devices, such as centrifuge safety cup with sealed rotor, must be used.
Use of BSCA BSC is the most important piece of
containment equipment but only
• if properly installed, • appropriate air velocity is maintained during
use, • proper procedures are used.
Use of biological safety cabinets
Facilities
–Ventilation–Temperature control–Sinks, eyewash, trashcans–Furniture and decontamination
Plan of a culture laboratory
Handling of contaminated laboratory supplies
• Glassware• Sputum containers• Applicator sticks, paper, pipettes• Positive and negative slides
Waste HandlingPotentially infectious materials must
be placed in a durable, leak proof container during collection, handling,
processing, storage, or transport within a facility.
No infected material should leave the laboratory except when it is properly packed for
transport to another laboratory.
Cleaning laboratory materials
Disinfection and Sterilization
A basic knowledge of disinfection and sterilization is crucial for biosafety in
the laboratory.
Laboratory equipment should be routinely decontaminated, as well as, after spills,
splashes, or other potential contamination.
Autoclaves
Boiling and burning
Safety Practices
• Pipetting• Microscopy
• Handwashing
Handwashing
Training
Initial on hireAnnual updates
Staff should be observed in their biosafety practices
Laboratory safety does not just happen.
The best defense against a laboratory accident is a well-thought-out plan to neutralize its
effects as quickly and effectively as possible.
• recognize that accidents can and will occur• formulate a plan of action• discuss ways to minimize and prevent
accidents
Spill ProceduresLaboratory accidents in the TB
laboratory result in possible formation of aerosols.
Spills involving infectious materials must be contained, decontaminated, and cleaned up by staff properly trained
and equipped to work with infectious material.
Emergency Procedures
• Puncture wounds, cuts and abrasions• Ingestion of potentially infectious material• Potentially infectious aerosol release (outside a
biological safety cabinet)• Broken containers and spilled infectious substances• Breakage of tubes containing potentially infectious
material in centrifuges not having sealable buckets• Breakage of tubes inside sealable buckets (safety
cups)
Incidents that may result in exposure to infectious materials must be immediately
evaluated and treated according to procedures described in the laboratory
biosafety manual. All such incidents must be reported to the laboratory supervisor. Medical evaluation, surveillance, and
treatment should be provided and appropriate records maintained.
Support Staff
The safe and optimum operation of a laboratory is dependent to a great extent
on the support staff, it is essential that such personnel are given
appropriate safety training.
THE MYCOBACTERIOLOGY LABORATORY IN NEED OF IMPROVEMENT
Physical facilities BSL-2 vs BSL-3
Laboratory Inspection
• Engineering controls• Administrative controls• Personal protective equipment• General Laboratory Safety
Risk Assessment
Will evaluate all procedures for risks related to aerosol generation and injury from
contaminated sharp objects (e.g., needle sticks) and develop a strategy for safe, step-by-
step manipulation of both specimens and cultures.
Why an assessment?Who should perform it?
When should it be done?What tools are used?
How should it be done?
Potential Hazards:
The following slides show some improper biosafety practices or containment safeguards
found in some TB laboratories.
Summary
Although the incidence of tuberculosis is higher in laboratory workers than for the general
population, the risk of becoming infected with M. tuberculosis in the laboratory can be
minimized through the use of the engineering controls, administrative procedures, and
specific work-place practices that are presented in these guidelines.
Questions?
Resources
• www.psmile.org• www.asm.org• www.clsi.org• www.cdc.gov• www.who.int./csr/resources/publications/biosafety/en/Biosafety7.pdf